Reversible site-specific tagging of enzymatically synthesized RNAs using aldehyde-hydrazine chemistry and protease-cleavable linkers.

Abstract:

:The investigation of RNA structure, dynamics and biological function often requires the site-specific incorporation of non-natural moieties. Here we describe the functionalization of RNA transcripts by aldehyde-hydrazine chemistry using a simple initiator nucleotide that carries an acetal-protected aldehyde function. This initiator nucleotide was efficiently incorporated into RNA, and the modified RNAs were quantitatively coupled to a peptide derivative displaying a hydrazine moiety at one end, a biotin tag at the other, and a trypsin-cleavable sequence in between. RNA conjugates could be easily isolated by affinity chromatography on streptavidin agarose and quantitatively cleaved off the support by trypsin treatment without detectable RNA degradation. The strategy described here may allow the incorporation of various new features into enzymatically synthesized RNA under mild conditions.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Pfander S,Fiammengo R,Kirin SI,Metzler-Nolte N,Jäschke A

doi

10.1093/nar/gkl1110

subject

Has Abstract

pub_date

2007-01-01 00:00:00

pages

e25

issue

4

eissn

0305-1048

issn

1362-4962

pii

gkl1110

journal_volume

35

pub_type

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